1. Field of the Invention
The present invention relates to a nucleic acid primer sequence, a kit, and a nucleic acid chain-immobilized carrier for detection of human papilloma virus and identification of its genotype, and a method of detecting human papilloma virus by using a nucleic acid amplification method.
2. Description of the Related Art
Human papilloma virus (HPV) infection was reported as a cause of uterine cervical cancer in the 1980's, and in particular, the relationship between cancer malignancy and HPV genotype is attracting attention. HPV is also considered to be the cause of cancers other than uterine cervical cancer such as cancers of the genital organs and oral mucosa, and there has been a demand for a rapid and accurate method of detecting HPV. Hitherto known were a method of detecting a malignant or benign genotype by using a DNA/RNA-recognizing antibody, and a method of amplifying a region containing a sequence characteristic to a genotype in polymerase chain reaction (PCR) and identifying the genotype finally by using a genotype-specific probe. However, the former method, which does not identify the genotype, is not applicable to the test for vaccine administration currently under development. Alternatively, the latter method of using the PCR method had disadvantages such as complicated procedure of pretreatment for example nucleic acid extraction, demand for a complex temperature-regulating device such as thermal cycler, and longer reaction period of two hours or more. In addition, the PCR method has a possibility that, if an incorrect complementary strand happens to be synthesized, the product may be used as a template in amplification, consequently leading to incorrect judgment. Actually, it is difficult to control specific amplification only with a difference of one nucleotide at the terminal of a primer.
For detection by using a DNA chip, gene products amplified by the PCR method are generally double-stranded chains. Thus, there emerged a problem that the complementary strands became competitors to the probe, lowering hybridization efficiency and detection sensitivity in the hybridization reaction with a probe. Accordingly, for example, a method of decomposing or separating the complementary strand is employed to make the target gene product into a single strand. However, these methods still have problems such as the higher cost and complicated procedure because of the use of enzymes or magnetic beads, and there exists a need for a new method replacing such conventional methods.